Characterization of the Felix domesticus (cat) glaucoma-associated protein myocilin

Michael P Fautsch, Anne M. Vrabel, Douglas H. Johnson

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The domestic cat (Felix domesticus) is a useful model for understanding the implications of long-term expression and function of normal and mutant myocilin. To better understand the role myocilin has in the cat eye, we isolated and characterized cat myocilin. Oligonucleotides designed against conserved nucleotide regions of myocilin mammalian orthologs were used to PCR amplify a partial cat myocilin cDNA clone. Rapid amplification of cDNA ends (5′ and 3′ RACE) was used to obtain full-length cat myocilin. The 2125 nucleotide cat myocilin cDNA contains a 490 amino acid open reading frame. Comparison of cat myocilin to human myocilin shows a 87% similarity, including conservation of the N-terminal leucine zipper, N-linked glycosylation site, C-terminal olfactomedin domain, and all five cysteine residues thought to be involved in disulfide bond formation. Expression in a transformed human trabecular cell line or in Crandall feline kidney cells showed cat myocilin was secreted from these cells, similar to human myocilin, suggesting cat myocilin contains a functional signal peptide sequence. In contrast, expression of cat myocilin containing a known human glaucoma-associated mutation (Y423H in cat; Y437H in human) was not secreted. Characterization of cat myocilin will enable long-term studies be performed in Felix domesticus to analyze changes to intraocular pressure and the aqueous outflow pathway following expression of myocilin and glaucoma causing mutations.

Original languageEnglish (US)
Pages (from-to)1037-1045
Number of pages9
JournalExperimental Eye Research
Volume82
Issue number6 SPEC. ISS.
DOIs
StatePublished - Jun 2006

Fingerprint

Glaucoma
Cats
Proteins
Complementary DNA
trabecular meshwork-induced glucocorticoid response protein
Nucleotides
Leucine Zippers
Mutation
Felidae
Protein Sorting Signals
Intraocular Pressure
Glycosylation
Oligonucleotides
Disulfides
Open Reading Frames
Cysteine
Clone Cells

Keywords

  • cat
  • cDNA
  • Felix domesticus
  • myocilin
  • TIGR

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

Characterization of the Felix domesticus (cat) glaucoma-associated protein myocilin. / Fautsch, Michael P; Vrabel, Anne M.; Johnson, Douglas H.

In: Experimental Eye Research, Vol. 82, No. 6 SPEC. ISS., 06.2006, p. 1037-1045.

Research output: Contribution to journalArticle

Fautsch, Michael P ; Vrabel, Anne M. ; Johnson, Douglas H. / Characterization of the Felix domesticus (cat) glaucoma-associated protein myocilin. In: Experimental Eye Research. 2006 ; Vol. 82, No. 6 SPEC. ISS. pp. 1037-1045.
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